[29] Monosaccharides attached to agarose

Barker, Robert; Chiang, Chao-Kuo; Trayer, Ian P.; Hill, Robert L.

doi:10.1016/s0076-6879(74)34032-3

Cited by 22 publications

(4 citation statements)

References 16 publications

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“…Synthesis of ATP-Agarose. The ATP-agarose used in the purification was synthesized by coupling the ligand [8-(6aminohexyl)amino-ATP] (Sigma) to cyanogen bromide (CNBr)-activated Sepharose-4B (Pharmacia) following a previously published procedure (Barker et al, 1974). All steps were performed at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

Heme Stoichiometry of Heterodimeric Soluble Guanylate Cyclase

Stone¹,

Marletta²

1995

Biochemistry

104

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The soluble form of guanylate cyclase (sGC) is to date the only definitive receptor for the novel signaling agent nitric oxide (.NO). .NO increases the Vmax of sGC by 100-200-fold, and it has been proposed that this activation occurs subsequent to the binding of .NO to a heme moiety on the enzyme. It has previously been demonstrated that the enzyme can be purified in a state containing as much as 1 heme per heterodimer. However, since the two subunits of the heterodimer display considerable homology, and the enzyme routinely loses heme upon purification, it has been unclear whether the native heme stoichiometry is 1 per heterodimer or 2 per heterodimer. Using a novel procedure, the enzyme has been purified to homogeneity from bovine lung in a state containing 1.52 +/- 0.10 equiv of heme per heterodimer, indicating that the native heme stoichiometry is 2 per heterodimer. The .NO-activated specific activity of this enzyme is increased by 50% over that of enzyme containing 1 heme per heterodimer and is the highest specific activity ever observed for sGC. Spectrally only one type of heme is observed, indicating that both hemes in the heterodimer are in similar environments. It is concluded that each subunit of the heterodimer binds 1 equiv of heme at a site conserved between the two subunits. Alignment of the nine published cDNA sequences for sGC indicates that the heme binding domain is the central portion of each subunit, corresponding to residues 213-370 in the bovine beta 1 sequence.

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Section: Methodsmentioning

confidence: 99%

Heme Stoichiometry of Heterodimeric Soluble Guanylate Cyclase

Stone¹,

Marletta²

1995

Biochemistry

104

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“…A new type of cluster ligand was prepared by attaching a number of different amino-containing glycosides (e.g., tris-LacAHT in Chart II) to the carboxyl groups of TV-Cbz-Asp (Scheme I). Coupling of the amino and the carboxyl group was accomplished by using either EEDQ as a coupling agent or the mixed anhydride method (Barker et al, 1974). Some examples of the structures and the corresponding abbreviated nomenclature of these cluster ligands are shown in Chart III.…”

Section: Methodsmentioning

confidence: 99%

New synthetic cluster ligands for galactose/N-acetylgalactosamine-specific lectin of mammalian liver

Lee¹,

Lin²,

Lee³

1984

Biochemistry

183

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Synthetic ligands containing up to six residues of nonreducing terminal galactose were prepared. The synthesis involved coupling of carboxyl groups of N-benzyloxy-carbonylaspartic acid or of N-benzyloxycarbonyltyrosyl-gamma-glutamylglutamic acid to the omega-amino group of the aglycon of a glycoside that contained up to three lactosyl residues. The benzyloxycarbonyl group was removed by hydrogenolysis before these ligands were tested as inhibitors to the binding of 125I-asialoorosomucoid to the galactose/N-acetylgalactosamine lectin, both soluble and on the surface of freshly isolated mammalian hepatocytes. Each addition of a galactosyl residue to an existing ligand structure invariably increased the binding affinity of such a ligand. However, at each level of galactose valency, the binding constant varied as much as 1000-fold depending on the structure of the ligand. At a given level of valency, the binding strength of a cluster ligand depended mainly on two factors: (1) the maximum spatial inter-galactose distances and (2) the flexibility of the arm connecting galactosyl residues and the branch points. It has been postulated that the three galactose-combining sites of the lectin are arranged in space at the vertexes of a triangle whose sides are 15, 22, and 25 A [Lee, Y. C., Townsend, R. R., Hardy, M. R., Lönngren, J., & Bock, K. (1984) in Biochemical and Biophysical Studies of Proteins and Nucleic Acids (Lo, T. B., Liu, T. Y., & Li, C. H., Eds.) pp 349-360, Elsevier, New York]. Ligands having inter-galactose distances shorter than these lengths were invariably poor ligands at their respective level of valency.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…Aminopropyl glycosides were prepared as described previously (29,31). Amino hexylglucoside was synthesized as described (32).…”

Section: Binding Of Siglec-8-ig To Aminoalkyl Glycosides Immobilized mentioning

confidence: 99%